Effect of a novel topical immunomodulator, S-28463, on keratinocyte cytokine gene expression and production

A new immunomodulating agent, imiquimod, has been reported to have antiviral and antitumor activities in animal models, S-28463 (4-amino-2-ethoxymethyl-alpha, alpha-dimethyl-1H-imidazo[4,5-c]quinoline-1-ethanol); an analog of imiquimod, has more potent antiviral activity in animals than imiquimod, It has also been shown to be more potent at inducing cytokines in human blood in vitro, However, its precise role as an immunomodulator in the skin has not been determined. We investigated the effect of S-28463 on human keratinocyte (KC) production of interferon-alpha (IFN-alpha) and other proinflammatory cytokines, including interleukin (IL)-1 alpha, IL-8, and tumor necrosis factor-alpha (TNF-alpha), Human KC were incubated with S-28463 at two concentrations (1 mu g/ml and 10 mu g/ml) for 6 h, Cytokine gene expression was analyzed by reverse-transcriptase PCR, In human KC, S-28463 stimulated significant increases in IFN-alpha mRNA at both concentrations. IL-1 alpha mRNA increased 1.4-fold at 10 mu g/ml. IL-8 mRNA was upregulated 2.5-fold at 10 mu g/ml. Twenty-four hours after treatment, IL-1 alpha, IL-8, and TNF-alpha protein mere increased, but IFN-alpha was below the level of detection. These results suggest that in the skin. S-28463-induced-IL-1 alpha; IL-8, and TNF-alpha production may be involved in the immunomodulating action of S-28463.